1. Make two primer probe sets
U5-Psi:
U5-Psi forward: TGTGTGCCCGTCTGTTGTGT
U5-Psi reverse: GAGTCCTGCGTCGAGAGAGC
U5-Psi probe: FAM-CAGTGGCGCCCGAACAGGGA-TAMARA

WPRE:
WPRE forward: CCGTTGTCAGGCAACGTG
WPRE reverse: AGCTGACAGGTGGTGGCAAT
WPRE probe: FAMTGCTGACGCAACCCCCACTGGT-TAMARA

2. Purify RNA from 10 ul virus stock using RNeasy.

3. Make serial dilution  from 1 ul RNA

4. Use ABI One-step RT-PCR Master mix with reverse transcriptase  for RT-qPCR.
Primer: 300 nM
Probe: 100 nM
50 ul vol
48 oC 30 min
95 oC 10 min
40 cycles of 95 oC for 15 sec and 60 oC for 1 min.